LOCUS       sequence-387031-        8685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 PhiC31 cassette with constitutive promoter: 
            pPP2AA3:PhiC31:t35S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8685)
  AUTHORS   Guiziou S, Maranas CJ, Chu JC, Nemhauser JL
  TITLE     An integrase toolbox to record gene-expression during plant 
            development.
  JOURNAL   Nat Commun. 2023 Apr 3;14(1):1844. doi: 10.1038/s41467-023-37607-5.
  PUBMED    37012288
REFERENCE   2  (bases 1 to 8685)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-04-3"; volume: "14(1):1844. doi"; pages: " 
            10.1038/s41467-023-37607-5"
COMMENT     Sequence Label: L1P46
FEATURES             Location/Qualifiers
     source          1..8685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    44..68
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(159..747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(239..258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2256..3071)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2256..3071)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2979..2998
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3063..3082
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3362..3797
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    3924..3946
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    3954..4130
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(4362..5387)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(5407..5752)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(5444..5464)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     5955..5977
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5969..5986
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5970..5986
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5993..6012
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5993..6011
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6594..6605
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6653..8470
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     polyA_signal    8508..8682
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 accacagaat caggggataa cgcaggaaag aacatgaagg ccttgacagg atatattggc
       61 gggtaaacta agtcgctgta tgtgtttgtt tgagatctca tgtgagcaaa aggccagcaa
      121 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      181 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      241 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      301 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      361 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      421 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      481 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      541 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      601 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaagaag agttggtagc
      661 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      721 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      781 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      841 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tactgagaga
      901 tcccctcata atttccccaa agcgtaacca tgtgtgaata aattttgagc tagtagggtt
      961 gcagccacga gtaagtcttc ccttgttatt gtgtagccag aatgccgcaa aacttccatg
     1021 cctaagcgaa ctgttgagag tacgtttcga tttctgactg tgttagcctg gaagtgcttg
     1081 tcccaacctt gtttctgagc atgaacgccc gcaagccaac atgttagttg aagcatcagg
     1141 gcgattagca gcatgatatc aaaacgctct gagctgctcg ttcggctatg gcgtaggcct
     1201 agtccgtagg caggactttt caagtctcgg aaggtttctt caatctgcat tcgcttcgaa
     1261 tagatattaa caagttgttt gggtgttcga atttcaacag gtaagttagt tgctagaacc
     1321 catggctcct ttgccgacgc tgagtagatt ttaggtgacg ggtggtgaca atgagtccgt
     1381 gtcgagcgct gattttttcg gcctttagag cgagatttat acaatagaat ttggcatgag
     1441 attggattgc ttttagtcag cctcttatag cctaaagtct ttgagtgact agatgacata
     1501 tcatgtaagt tgctgatagg tttccagttt tccgctccta ggtctgcata ttgtactttt
     1561 cctcttactc gacttaacca gtaccaaccc agcttctcaa cggatttata ccatggcact
     1621 ttaaagccag catcactgac aatgagcggt gtggtgttac tcggtagaat gctcgcaagg
     1681 tcggctagaa attggtcatg agctttcttt gaacattgct ctgaaagcgg gaacgctttc
     1741 tcataaagag taacagaacg accgtgtagt gcgactgaag ctcgcaatac cataagtcgt
     1801 ttttgctcac gaatatcaga ccagtcaaca agtacaatgg gcatcgtatt gcccgaacag
     1861 ataaagctag catgccaacg gtatacagcg agtcgctctt tgtggaggtg acgattacct
     1921 aacaatcggt cgattcgttt gatgttatgt tttgttctcg ctttggttgg caggttacgg
     1981 ccaagttcgg taagagtgag agttttacag tcaagtaatg cgtggcaagc caacgttaag
     2041 ctgttgagtc gttttaagtg taattcgggg cagaattggt aaagagagtc gtgtaaaata
     2101 tcgagttcgc acatcttgtt gtctgattat tgatttttcg cgaaaccatt tgatcatatg
     2161 acaagatgtg tatccacctt aacttaatga tttttaccaa aatcattagg ggattcatca
     2221 gtctaaagta tatatgtgta acattggtct agtgattaga aaaactcatc gagcatcaaa
     2281 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2341 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2401 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2461 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     2521 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2581 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2641 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2701 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2761 ttccctggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2821 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2881 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2941 tacaatcggt agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3001 tataaatcag catccatgtt ggaatttaat cgcggccttg agcaagacgt ttcccgttga
     3061 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     3121 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     3181 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
     3241 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
     3301 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggcgatc
     3361 cccatccaac agcccgccgt cgagcgggct tttttatccc cggaagcctg tggatagagg
     3421 gtagttatcc acgtgaaacc gctaatgccc cgcaaagcct tgattcacgg ggctttccgg
     3481 cccgctccaa aaactatcca cgtgaaatcg ctaatcaggg tacgtgaaat cgctaatcgg
     3541 agtacgtgaa atcgctaata aggtcacgtg aaatcgctaa tcaaaaaggc acgtgagaac
     3601 gctaatagcc ctttcagatc aacagcttgc aaacacccct cgctccggca agtagttaca
     3661 gcaagtagta tgttcaatta gcttttcaat tatgaatata tatatcaatt attggtcgcc
     3721 cttggcttgt ggacaatgcg ctacgcgcac cggctccgcc cgtggacaac cgcaagcggt
     3781 tgcccaccgt cgagcgcctt tgcccacaac ccggcggccg gccgcaacag atcgttttat
     3841 aaattttttt ttttgaaaaa gaaaaagccc gaaaggcggc aacctctcgg gcttctggat
     3901 ttccgatccc cggaattaga tcttggcagg atatattgtg gtgtaacgtt atcgatctgg
     3961 attttagtac tggattttgg ttttaggaat tagaaatttt attgatagaa gtattttaca
     4021 aatacaaata catactaagg gtttcttata tgctcaacac atgagcgaaa ccctataaga
     4081 accctaattt cccttatcgg gaaactactc acacattatt tatggagaaa atagagagag
     4141 atagatttgt agagagagac tggtgatttc agcgtaccga attaattcct gaatcactgc
     4201 gaccggccct cccgcgaccc agccgagcga gcttagcgaa ctgtggacga gaactgtgcc
     4261 accaagcgta aggccgttct ctcgcatttg ccttgctagg ctcgcgcgag ttgctggctg
     4321 aggcgttctc gaaatcagct cttgttcggt cggcatctac tctattcctt tgccctcgga
     4381 cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag
     4441 acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg
     4501 attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc
     4561 tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccttgg cgatcctgca
     4621 agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc
     4681 ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag
     4741 tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg
     4801 tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc
     4861 tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga
     4921 tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg
     4981 aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatagcatc catagcctcc
     5041 gcgaccggct gaagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc
     5101 tgtgcacggc gggagatgca ataggtcagg ctctcgctga actccccaat gtcaagcact
     5161 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa
     5221 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa
     5281 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg
     5341 atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatagg ggggatcagc
     5401 ttgggctgtc ctctccaaat gaaatgaact tccttatata gaggaagggt cttgcgaagg
     5461 atagtgggat tgtgcgtcat cccttacgtc agtggagatg tcacatcaat ccacttgctt
     5521 tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg ggggtccatc
     5581 tttgggacca ctgtcggcag aggcatcttg aatgatagcc tttcctttat cgcaatgatg
     5641 gcatttgtag gagccacctt ccttttctac tgtcctttcg atgaagtgac agatagctgg
     5701 gcaatggaat ccgaggaggt ttcccgaaat taccctttgt tgaaaagtct caatagccct
     5761 ttggtcttct gagactgtat ctttgacatt tttggagtag gggtacgata acattaacgc
     5821 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5881 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     5941 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     6001 ctcactatag ggcttggagg aggaagaaga agagagttac tgagaagctt cctttatacg
     6061 agagagatat taaacgaagt cgtatttgtt ggatatatgg gcctttatgt attatacata
     6121 agcccattta gttcttatct ttgacaggcc caattatatg gacacagctt gaagatggtt
     6181 tagctgctgc gaaagacgag ctccggtctg atttctcgtt cttctgattg atagatcgct
     6241 cggaacttgg aaagcagcgt aatcggtaaa ttctcgatca tttcttcaat ttcgaaacat
     6301 tttccaattg tatattgaga atctcaaatc gaattattgt ttagatcttc tctctttatt
     6361 agtctaagaa tggtaatagt ttctctctag attgggacgg cacaaagttt cagttctcca
     6421 ttctaatcat tttttagagg ctaatgggtc acctttgaat tatctcagag gcggaattga
     6481 gtttaaagtt tgtaacttta tcgtcttttg tattgatggt gtctatggca aaggtgatta
     6541 actgcaaagg tatgcattgt gttttattct gactttgtgt acatttgtgt ttgattgaag
     6601 gtagggagtg gcaaaggtga ttaactgcaa aggatttgag ttttggtgag gaatggacac
     6661 gtatgccggc gcttatgata gacaatcaag agaaagagag aactcttctg ccgcctctcc
     6721 agctacacag agatctgcca acgaggataa agctgctgat ttacaaagag aggtagagag
     6781 agatgggggt aggtttagat ttgtgggtca tttctctgag gctcctggta catcagcctt
     6841 tggtactgct gaacggcctg aatttgaaag aatattaaat gagtgtcggg ccggtcgact
     6901 aaatatgata atagtttatg atgtatctag gttcagtaga ctcaaagtta tggatgcaat
     6961 tccgatcgtg agcgagctac ttgcactcgg tgttactatc gtaagtacac aagaaggtgt
     7021 attcaggcag ggtaatgtga tggaccttat acaccttatt atgcgactcg acgctagcca
     7081 taaagagtca tcactgaaga gtgctaaaat cttggacacc aagaacctgc aaagggagct
     7141 tggaggctat gtaggaggaa aagctccgta tggtttcgaa ctcgtgagtg agacaaaaga
     7201 gattaccaga aatggtagga tggtgaacgt ggttattaat aagctagctc atagcacaac
     7261 ccctctcaca gggccttttg agttcgaacc tgacgttatt agatggtggt ggagagagat
     7321 caaaactcac aaacatcttc catttaagcc aggtagccag gctgctatcc accctggatc
     7381 tatcactggg ctttgtaaaa ggatggatgc tgacgcagtg ccaactcgag gggaaacaat
     7441 cggtaagaag actgcatctt ccgcttggga tcccgctaca gttatgagaa tccttcggga
     7501 tccgcgtatt gctggattcg ccgcagaagt catttataag aaaaaacctg atggtactcc
     7561 tactacaaaa atcgaaggtt ataggatcca aagggatcct attactttgc gtccagttga
     7621 attagactgt ggaccgatca tcgaacctgc cgagtggtac gagttgcagg cctggcttga
     7681 tggtagaggg cgtggaaagg gtttatctcg gggacaggca attttgtcag caatggataa
     7741 gctttattgt gagtgtgggg ctgttatgac atcaaaacgt ggagaagagt caatcaagga
     7801 ttcttacagg tgcagacgtc gtaaagtggt ggacccttct gctccaggac agcatgaggg
     7861 tacatgcaac gtttctatgg ctgctcttga taaatttgtg gccgaacgta ttttcaacaa
     7921 gattagacac gctgaagggg atgaggaaac tttagctctt ctttgggagg ctgcccggcg
     7981 ttttggtaag ctaacagagg caccagaaaa aagtggcgaa agagctaatt tggtagccga
     8041 aagagcagac gccttgaatg cattggaaga actttacgaa gatagggctg ccggagcata
     8101 tgatggacca gttggtcgga agcatttcag gaagcaacaa gctgcactaa ctttgagaca
     8161 gcaaggcgca gaggagcgac tcgcagagct tgaggccgcc gaggccccaa agcttccact
     8221 cgatcaatgg tttcctgagg atgccgacgc tgatcctaca ggtccaaaga gctggtgggg
     8281 aagagctagc gtagacgata agagagtgtt tgtgggtcta tttgttgaca agattgtagt
     8341 tactaagtcc actacgggtc gaggacaggg cactcccatc gaaaagagag ccagtattac
     8401 ctgggcaaag cctccaaccg atgatgatga agacgatgca caagatggta ccgaggatgt
     8461 ggcagcataa taagcttctc tagctagagt cgatcgacaa gctcgagttt ctccataata
     8521 atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg ctcatgtgtt
     8581 gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat
     8641 ttctaattcc taaaaccaaa atccagtact aaaatccaga tcgct
//
